Animal Tissue Culture Basics

Questions and Answers

Which of the following substances is classified as a carcinogen?

Streptomycin

Hygromycin (correct)

Gentamycin

Thapsigargin

What is cellular senescence?

The process of cells undergoing mutation

The phenomenon where cells lose the ability to divide (correct)

The ability of cells to divide indefinitely

The method of isolating cells from tissues

What method is used to release mononuclear cells from soft tissues for culture?

Mechanical disruption of tissue

Using a centrifuge to separate components

Placing tissue pieces in saline solution

Enzymatic digestion with collagenase (correct)

Which of the following statements about primary cell cultures is accurate?

They have a limited lifespan with a defined number of population doublings. Signup and view all the answers

What defines an established or immortalized cell line?

Cells that can proliferate indefinitely. Signup and view all the answers

What is a primary cell culture primarily derived from?

Dispersed cells from the original tissue Signup and view all the answers

What characterizes an established or continuous cell line?

It has infinite growth potential due to transformation. Signup and view all the answers

What is the purpose of a feeder layer in cell culture?

To serve as a substrate for other cells to grow on. Signup and view all the answers

Which of the following is a common application of tissue culture?

Producing monoclonal antibodies. Signup and view all the answers

What type of substrate may be used in cell culture?

Solid, semisolid, or liquid Signup and view all the answers

What does passage number refer to in cell culture?

The number of times cells have been transferred to a new vessel. Signup and view all the answers

Which factor is NOT typically regulated in the culture environment of cells?

Color of the culture medium Signup and view all the answers

What is one feature of the media choice in cell culture?

It is determined empirically for specific cell types. Signup and view all the answers

What is the primary purpose of cell/tissue culture?

To propagate cells outside the organism Signup and view all the answers

Which of the following statements is true about tissue culture?

Tissue culture results in clones with the same genotype Signup and view all the answers

What significant advancement did Enders, Weller, and Robbins achieve in 1948?

Proved poliomyelitis virus can be grown in vitro without nerve tissue Signup and view all the answers

What was the main contribution of George Gey in 1952?

Establishing the first human cell line from cervical carcinoma Signup and view all the answers

What is the primary challenge associated with growing large complex proteins in E. coli?

E. coli does not provide proper glycosylation Signup and view all the answers

Which technique allows for the maintenance of the structural integrity of tissue in culture?

Organ culture Signup and view all the answers

Which of the following describes a significant aspect of the 1955 advances in cell culture media?

Provided consistency and reduced chances of contamination Signup and view all the answers

What is a characteristic of normal human diploid cells as described by Hayflick and Moorhead?

They exhibit a finite life span Signup and view all the answers

What is a major difference between transformed cells and primary cells in cell culture?

Transformed cells can grow without cell-cell contact. Signup and view all the answers

Which of the following is NOT a limitation of cell culture?

Total inability to grow any cells Signup and view all the answers

What is a significant feature that cell culture lacks when compared to normal tissue?

Blood circulation Signup and view all the answers

Which characteristic of transformed cells can lead to variability in experimental outcomes?

High adaptability to various conditions Signup and view all the answers

What might be a consequence of using hormones in high concentrations in cell culture?

Alteration in cell types Signup and view all the answers

Which factor contributes to the genetic instability of cells in culture?

Heterogeneity of cell populations Signup and view all the answers

What is a critical environmental control issue when conducting cell culture experiments?

Maintaining a specific pH level Signup and view all the answers

Why is cell contact significant for primary cells in culture?

It affects genetic expression. Signup and view all the answers

What characteristic of tissue culture allows for experiments to be conducted without the variations seen in animal studies?

Control of the growth environment Signup and view all the answers

Which of the following advantages is a characteristic of tissue culture?

Ability to expose cultures to defined concentrations of reagents Signup and view all the answers

One of the limitations of tissue culture is that it lacks the physiological complexity of living organisms. What specifically is this limitation related to?

Absence of organized tissues Signup and view all the answers

How can tissue cultures aid in reducing ethical concerns related to research?

By minimizing animal experimentation Signup and view all the answers

What is a common disadvantage of tissue culture that researchers must address?

Dedifferentiation and potential loss of cellular mechanisms Signup and view all the answers

Which factor can be controlled in the physico-chemical environment of tissue culture?

Osmolarity and pH levels Signup and view all the answers

Which aspect of tissue culture helps ensure that a consistent lineage of cells is maintained over generations?

Selective media and cloning availability Signup and view all the answers

What is a characteristic challenge faced by researchers when working with tissue cultures?

Learning and maintaining aseptic techniques Signup and view all the answers

Study Notes

Overview of Cell/Tissue Culture

Cell culture is the propagation of cells outside an organism, involving both plant and animal cells.
Tissue culture produces clones with identical genotypes unless mutations occur during culture.
Environmental conditions in culture must replicate the organism's conditions: temperature, humidity, nutrition, and contamination-free.

History of Cell Culture

1801: Bichat names human body tissues.
1880: Roux maintains embryonic chicken tissue in saline.
1907: Harrison demonstrates in vitro growth of living animal tissue from a tadpole nerve cord.
1911-1912: Burrows and Carrel grow explants from adult animals, employing aseptic techniques.
1948: Enders, Weller, and Robbins grow polio virus in vitro using monkey kidney cells.
1952: George Gey establishes the HeLa cell line from human cervical carcinoma.
1961: Hayflick and Moorhead identify finite lifespan of normal human diploid cells.

Terminology

Organ culture: Three-dimensional culture retaining tissue features in vivo.
Cell culture: Single cells derived from dispersed tissue cells.
Primary cell culture: Directly derived from explants with a limited lifespan.
Clone: A population derived from a single original cell.
Sub-culture: Transplantation of cells to new vessels.
Established cell lines: Immortalized cell cultures, often tumor-derived, like CHO and SH-SY-5Y cells.
Passage number: Successive sub-cultures from the primary culture.

Applications of Cell Culture

Research: Understand cell functions and toxicology.
Tissue engineering: Cultivation of skin, liver, bone, and cartilage.
Production of biologicals: Create monoclonal antibodies, hormones, enzymes, and vaccines.

Culture Environment

Substrates can be solid (glass, plastic), semisolid (agar, collagen gel), or liquid (suspension cultures).
Feeder layers support growth by preventing proliferation.
Gas phase: Standard CO2 and O2 levels must be maintained to optimize cell growth.
Media must provide nutrients and be cell-type specific, ensuring sterility and isotonic conditions.

Advantages of Tissue Culture

Enables cell behaviour study without variations from whole organisms.
Controlled growth environments ensure sample uniformity.
Cells retain characteristics over generations, providing reproducibility.
Large quantities of cells can be produced.
Reduces ethical concerns related to animal experimentation.

Shortcomings and Limitations

In vitro culture lacks true physiological conditions and the complex interactions present in vivo.
Cells can undergo dedifferentiation, leading to loss of original features.
Standardized techniques are required for successful cell maintenance.
Limited material quantity and costly consumables can be a barrier.

Missing Features in Cell Culture

Unlike in vivo conditions, blood circulation, tissue organization, and cellular interaction are often absent in cultures.
Hormones are typically added in high concentrations, unlike natural physiological levels.

Safety in Cell Culture

Be aware of hazardous substances: carcinogens, teratogens, and mutagens.
Common examples include gentamycin (possible teratogen), hygromycin (possible carcinogen), and streptomycin (mutagen).

Types of Tissue Culture

Cells isolated from tissues can be cultured using enzymatic digestion techniques or explant cultures.
Primary cells have a limited lifespan, undergoing senescence after several divisions.
Established or immortalized cell lines can proliferate indefinitely, often used for research and diagnostics.

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Description

Explore the fundamentals of animal tissue culture, a technique for growing and maintaining cells outside their natural environment. This quiz covers the principles of cell and tissue culture, including the propagation of genetically identical clones. Test your knowledge on the methods and applications of this crucial biological technique.